Abstract:
To obtain monoclonal antibodies (McAbs) against E7 oncoprotein of human papillomavirus type 58 (HPV-58),lactose induction was applied to express GST-HPV58E7 fusion protein in recombinant bacterial strain.Using simply GST affinity chromatographic separation,the target protein was purified and used as a novel antigen to produce antibodies.After mouse immunization,somatic hybridization,cloned culture,positive hybridoma scanning,ascites preparation,rProtein,and affinity purification,6 strains of purified and high titer anti-HPV58E7 McAbs were prepared.Further studies used ELISA,Western-blot and immunofluorescence,and proved these obtained antibodies could specifically react with HPV-58 E7 which was expressed either in prokaryotic or eukaryotic cells.Moreover,another indirect ELISA also revealed that these 6 strains of McAbs were IgG2a κ type antibodies and bound with the same epitope in entire sequence of HPV-58 E7 oncoprotein. In conclusion,the prepared McAbs provides the basis for tumor biology research and the development of related diagnostic reagents which are targeting HPV-58 E7,and these studies are very important to effective screening,diagnosis,prevention and treatment of cervical cancer caused by HR-HPV detect in our country.